Interaction of poly(rC)-binding protein 2 domains KH1 and KH3 with coxsackievirus RNA.
Recombinant hnRNP K-homology (KH) domains 1 and 3 of the poly(rC)-binding protein (PCBP) 2 were purified and assayed for interaction with coxsackievirus B3 RNA in electrophoretic mobility shift assays using in vitro transcribed RNAs which represent signal structures of the 5'-nontranslated region. KH domains 1 and 3 interact with the extended cloverleaf RNA and domain IV RNA of the internal ribosome entry site (IRES). KH1 but not KH3 interacts with subdomain IV/C RNA, whereas KH3 interacts with subdomain IV/B. All in vitro results are consistent with yeast three-hybrid experiments performed in parallel. The data demonstrate interaction of isolated PCBP2 KH1 and KH3 domains to four distinct target sites within the 5'-nontranslated region of the CVB3 genomic RNA.
Zell
Roland
R
Institute for Virology and Antiviral Therapy, Friedrich Schiller University, Hans-Knöll-Str. 2, D-07745 Jena, Germany. Electronic address: Roland.Zell@med.uni-jena.de.
Ihle
Yvonne
Y
Biomolecular NMR Spectroscopy, Leibniz Institute for Age Research, Fritz Lipmann Institute, Beutenbergstr. 11, D-07745 Jena, Germany.
Effenberger
Madlen
M
Institute for Virology and Antiviral Therapy, Friedrich Schiller University, Hans-Knöll-Str. 2, D-07745 Jena, Germany.
Seitz
Simone
S
Institute for Virology and Antiviral Therapy, Friedrich Schiller University, Hans-Knöll-Str. 2, D-07745 Jena, Germany.
Wutzler
Peter
P
Institute for Virology and Antiviral Therapy, Friedrich Schiller University, Hans-Knöll-Str. 2, D-07745 Jena, Germany.
Görlach
Matthias
M
Biomolecular NMR Spectroscopy, Leibniz Institute for Age Research, Fritz Lipmann Institute, Beutenbergstr. 11, D-07745 Jena, Germany.
D024202:Electrophoretic Mobility Shift Assay; D004770:Enterovirus / Q000378:metabolism*; D034441:Heterogeneous-Nuclear Ribonucleoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009690:Nucleic Acid Conformation; D017434:Protein Structure, Tertiary; D022661:RNA, Untranslated / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism
D034441:Heterogeneous-Nuclear Ribonucleoproteins; C501903:PCBP2 protein, human; D022661:RNA, Untranslated; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins